Exercise No.

3
Nuclear and Cell Division

Introduction:

In meristematic tissues (plant tissues in which cells multiply), cells go through an orderly
sequence of events known as the cell cycle. The cycle is usually divided into interphase and
mitosis; mitosis is usually accompanied by cytokinesis (cell division). A clear understanding of
the process of mitosis is fundamental to an understanding of growth and reproduction, and helps
in the comprehension of the important process of meiosis. All living organisms undergo mitosis
in at least some stages of their life cycles. Plants and animals differ in some details of the
process, but not in the overall scheme.

Objectives:
1. Understand and know what takes place in each of the phases of mitosis
2. Understand the nature, structure, or function of spindle fibers, poles, equator, chromatids,
cell plate, and phragmoplast

Materials:
Slides of mitosis in onion (Allium) root tip
Two fresh onions with growing roots
One dropper bottle of acetocarmine stain
Models/charts of cells undergoing mitosis

Procedure:

Mitosis in Plant Cells:

Examine the Allium root tip slide under the microscope. The end of the root is protected
by a root cap consisting of cells that are irregular in shape. Immediately behind the root cap is a
meristematic region (apical meristem) where cells actively divide. Above the meristematic
region, in the region of elongation, the cells increase in length without dividing. To find cell in
the various stages of mitosis, you will have to confine your examination of the slide to the
meristematic region.

Interphase – Look for cells with distinct nuclei, each containing one, two or more nucleoli (which
appear as more or less spherical miniature nuclei within a nucleus. Most of the cells will be in
interphase. The fine, wispy, lightly stained material is called chromatin.

Prophase – In prophase, the nuclear envelope (“casing” of the nucleus) and nucleoli disappear,
the chromosome strands coil tightly, becoming shorter and thicker, and both strands of each
chromosome become distinguishable.

Metaphase – In metaphase, the centromeres of the chromosomes are aligned at the equator (an
invisible plate in the center) of the cell. A top-shaped spindle consisting of numerous spindle
fibers, become fully developed. The spindle fibers extend in arcs between two invisible poles
located toward opposite ends of the cell. Each chromosome has a single spindle fiber attached to its
centromere, but most of the spindle fibers are directly between poles.

Anaphase – In anaphase, the two strands of each chromosome (chromatids) separate longitudinally at
their centromeres. One strand of each pair migrates to a pole, and the other strand of each pair
simultaneously migrates to the opposite pole.

Telophase – In telophase, the tight coiling of the chromatids – now called chromosomes again – relaxes,
and the strands become longer and thinner; a new nuclear envelop develops around each group, and
nucleoli reappear. The spindle gradually disappears, and a set of shorter fibers, comprising the
phragmoplast, develops at right angles to the spindle between the two new nuclei. While this is taking
place, dictyosomes in the cytoplasm produces vesicles containing raw materials for cell walls and
membranes. Some of these vesicles are channeled to the center of the spindle (equator) by the remaining
fibers. As the vesicles accumulate, they fuse together, forming a double membrane called the cell plate.
The cell plate grows outward from the center until it contacts and unites with the plasma membrane.
Cellulose is then deposited on the membranes, resulting in the new cell walls. At the same time, pectin is
added between the membranes, creating a middle lamella that is shared at the end of telophase by what
has become two new daughter cells.

Optional Exercise

Observe the various stages of mitosis in freshly grown onion root tips. Place a drop of
acetocarmine (a mixture of acetic acid and a red stain) on a clean glass slide. Obtain a fresh onion root
and remove about 0.5 cm of the tapered tip end, which should be placed in the acetocarmine drop and
allowed to sit for about 5 minutes to soften the tissue. Then with a probe and a single-edged razor blade,
mash the root tip thoroughly (a second drop of acetocarmine may be added). Add a coverslip, then
observe under the microscope using HPO. How many stages of mitosis can you find in the stained cells?

Exercise to be Submitted

Label the four phases of mitosis (prophase, metaphase, anaphase, telophase) from cells in the
meristematic region of an onion root tip. Include the following: chromosomes, cell wall, spindle fibers,
centromere, cytoplasm, cell plate, nucleus, nucleolus, nuclear envelope, and phragmoplast. You do not
need to label each item more than once.